AmpC酶抑制剂筛选方法的建立及136种中药材活性评价  

Establishment of screening method for AmpC enzyme inhibitors and activity evaluation of 136 Chinese medicinal materials

作　　者：罗满平 孟瑛瑛 李德民 王康旭 李媚媛 张新国[1] 王宗元 LUO Man-ping;MENG Ying-ying;LI De-min;WANG Kang-xu;LI Mei-yuan;ZHANG Xin-guo;WANG Zong-yuan(Key Laboratory of Screening and Deep Processing in New Tibetan Medicine of Gansu Province,School of Life Science and Engineering,Lanzhou University of Technology,Lanzhou 730050,China;Lanzhou Animal Disease Prevention and Control Center,Lanzhou 730050,China)

机构地区：[1]兰州理工大学生命科学与工程学院,甘肃省中藏药筛选评价及深加工重点实验室,甘肃兰州730050 [2]兰州市动物疫病预防控制中心,甘肃兰州730050

出　　处：《中草药》2023年第19期6351-6361,共11页Chinese Traditional and Herbal Drugs

基　　金：国家自然基金资助项目(32160021);甘肃省重点研发计划项目(21YF5FA084,22YF7NA176);温州市科技局项目(2022Y0883)。

摘　　要：目的建立一种AmpC酶抑制剂的快速筛选方法,为治疗由耐药菌传播引起的感染提供筛选依据。方法以阿维巴坦钠为阳性对照,采用紫外分光光度法检测反应体系的吸光度变化,通过6种不同底物的筛选比较及反应条件的优化,并对136种中药提取物进行筛选评价。结果最佳体系为波长489 nm、底物质量浓度0.1 mg/mL、酶浓度400 U/mL、温度30℃、反应时间300 s。136种中药水提物中有12种中药水提物对AmpC酶的抑制率大于70%,其中桑寄生、赤芍、红景天、千斤拔、石见穿5种中药水提物的抑制率大于80%;添加桑寄生可明显提高青霉素的协同抗菌效应,最低抑菌浓度(minimal inhibit concentration,MIC)由2 mg/mL降低至0.8 mg/mL。结论建立的AmpC酶抑制剂筛选方法快速、高效、可信度高,中药是抗耐药菌感染治疗的重要资源。Objective To establish a rapid screening method for AmpC enzyme inhibitors to provide a basis for screening drugs in treatment of infections caused by spread of drug-resistant bacteria.Methods Avibactam sodium was used as a positive control,and the absorbance changes of reaction system were detected by UV spectrophotometry.Six different substrates were compared for screening,and the reaction conditions were optimized.A total of 136 Chinese medicinal materials extracts were screened and evaluated.Results The optimal system was wavelength 489 nm,substrate concentration 0.1 mg/mL,enzyme concentration 400 U/mL,temperature 30℃,and reaction time 300 s.Among the 136 aqueous extracts of Chinese medicinal materials,12 extracts showed AmpC enzyme inhibitory activities greater than 70%,and five extracts including Taxillus chinensis,Paeonia veitchii,Rhodiola crenulata,Flemingia philippinensis and Salvia chinensis had inhibitory activities greater than 80%,and the addition of T.chinensis significantly improved the synergistic antibacterial effect of penicillin,minimal inhibit concentration(MIC)was changed from 2 mg/mL to 0.8 mg/mL.Conclusion The established screening method for AmpC enzyme inhibitors is rapid,efficient and highly reliable,and traditional Chinese medicines are important resources for the treatment of infections caused by drug-resistant bacterial.

关 键 词：耐药菌 AMPC酶 抑制剂 筛选方法 桑寄生 赤芍 红景天 千斤拔 石见穿 

分 类 号：R285.51[医药卫生—中药学]